人外周血淋巴细胞pig3和gdf15基因表达变化作为辐射生物剂量计的研究
发布时间：2019-07-03 16:20
【摘要】：目的： 随着人类核活动的日益增多,核与辐射的意外暴露以及核恐怖事件发生的可能性也较以往有所增大。在大人群受到照射的情况下,传统的生物剂量估算方法双着丝粒和着丝粒环畸变分析不能满足快速生物剂量估算的需求,因此需要建立一种快速、高通量的生物标志物,以应对大批量人群辐射暴露情况下的生物剂量估算。因此本研究试图使用目前较为先进的Real-time PCR、ELISA等技术,对电离辐射诱导的人外周血淋巴细胞中pig3和gdfl5基因表达进行定量测定,并对其变化规律进行分析,这将有助于电离辐射的分子水平生物效应及其机制的阐明,为辐射损伤研究提供新的生物标志物,也将为发现更加有效、快速、高通量的生物剂量计积累理论依据。 方法： 1、在知情同意情况下,收集73名正常人外周血样品。利用Taqman MGB探针法绝对定量分析正常人外周血淋巴细胞中pig3和gdf15基因表达水平的本底值。 2、利用Taqman MGB探针法和ELISA技术分析受照射人外周血淋巴细胞中pig3和gdf15基因在照后多个时间点(4h、8h、12h、24h、48h和72h)转录水平和翻译水平的剂量效应关系,剂量为0、1、3、5和8Gy,最终通过统计学方法拟合剂量效应曲线；并确定时间响应范围。 3、通过分析受到不同剂量(OGy、2Gy、4Gy.6Gy)60Co γ射线照射后SD大鼠离体和整体外周血中gdf15基因表达变化,验证两种基因整体与离体效应是否一致,探索两种基因作为辐射生物剂量计应用的准确性。 结果： 1、在转录水平,正常人外周血pig3和gdf15基因表达的本底拷贝数在不同性别和不同年龄组之间没有统计学差异。 2.0～8Gy60Coγ射线照射后8h～48h人外周血淋巴细胞pig3和gdf15基因表达水平均呈现良好的剂量效应关系,并可拟合成为R2值较高的直线；照射后72h出现了随剂量增加,pig3和gdf15基因表达先上调后下调的趋势。 3、在翻译水平,0～8Gy60Coγ射线照射后48h和72h,在一定的剂量范围内,人外周血淋巴细胞PIG3蛋白表达呈现一定的剂量效应关系,可以拟合成为R2值较高的直线；其他时间点没有明显的剂量效应关系。 结论： 1、止常人外周血pig3和gdf15基因mRNA本底水平不受供血者性别和年龄的影响,这是两种基因表达水平成为辐射生物剂量计的基础。 2、在mRNA水平,离体照射的人外周血pig3和gdf15基因表达在0～8Gy剂量照射后8h～48h均表现出了良好的剂量效应关系,证实了两种基因表达变化具备了成为生物剂量计的基本条件。 3、PIG3蛋白水平在0～5Gy剂量照射后48h和72h具有良好的剂量效应关系。
[Abstract]:Purpose: With the increasing number of human nuclear activities, the accidental exposure of nuclear and radiation, as well as the possibility of nuclear terrorism, has also increased in the past Large. In the case of large population exposure, the traditional biological dose estimation method, the double centromere and the centromere ring distortion analysis, cannot meet the requirement of rapid biological dose estimation, therefore, a rapid and high-flux biological marker needs to be established. To deal with the estimation of the biological dose in the case of exposure to radiation in a large number of people In this study, we tried to measure the expression of pg3 and gdfl5 in human peripheral blood lymphocytes induced by ionizing radiation by using the technology of real-time PCR, ELISA and so on. This will help the molecular level biological effect of ionizing radiation and its mechanism, provide a new biological marker for the research of radiation damage, and will also find a more effective, rapid and high-flux biological dosimeter accumulation theory. It was reported. Method:1. Under the condition of informed consent,73 normal persons were collected. quantitative analysis of the expression of pg3 and gdf15 in peripheral blood lymphocytes from peripheral blood samples by using the Taqman MGB probe method the level of the pg3 and gdf15 genes in the peripheral blood lymphocytes of the irradiated human were analyzed by using the Taqman MGB probe method and the ELISA technique, and the dose effect relationship between the transcription level and the translation level of the human peripheral blood lymphocytes after the irradiation (4h, 8h, 12h, 24h, 48h and 72h) was analyzed, and the dose was 0,1, 3,5 and 8 Gy, and the dose response curves were finally fitted by a statistical method; and 3. By analyzing the expression of gdf15 gene in the isolated and whole peripheral blood of SD rats exposed to different doses (OGy, 2Gy, 4Gy. 6Gy), the expression of gdf15 gene in the isolated and whole peripheral blood of SD rats was changed. dosimeter The results were as follows:1. The background copy number of the expression of the pg3 and gdf15 genes in the peripheral blood of the normal human is different from the different sex and not in the normal human. There was no statistical difference between the same age groups. The expression levels of the pg3 and gdf15 gene in the peripheral blood lymphocytes of the human peripheral blood lymphocytes from 8 h to 48 hours after the irradiation of 2.0 to 8 Gy 60Co showed a good dose-effect relationship and can be fitted into a straight line with a higher R2 value. The dose-increasing, the pige3 and the gdf occurred at 72 h after the irradiation. The expression of PIG3 protein in peripheral blood of human peripheral blood showed a dose-effect relationship at 48 h and 72 h after the level of translation, and the expression of PIG3 protein in human peripheral blood lymphocytes could be fit into a straight line with high R2 value. ; others Conclusion:1. The level of the normal human peripheral blood pige3 and gdf15 gene mRNA is not affected by the sex and age of the donor. The expression level of the two genes was the basis of the radiation biological dosimeter.2. The expression of the human peripheral blood pige3 and gdf15 in the in-vitro irradiation at the mRNA level showed a good dose-effect relationship between 8 and 48 hours after the dose of 0-8Gy. The expression of the two genes is the basic condition of the biological dosimeter.3, the level of the PIG3 protein is 0-5.
【学位授予单位】：中国疾病预防控制中心
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R142
本文编号：2509519
[Abstract]:Purpose: With the increasing number of human nuclear activities, the accidental exposure of nuclear and radiation, as well as the possibility of nuclear terrorism, has also increased in the past Large. In the case of large population exposure, the traditional biological dose estimation method, the double centromere and the centromere ring distortion analysis, cannot meet the requirement of rapid biological dose estimation, therefore, a rapid and high-flux biological marker needs to be established. To deal with the estimation of the biological dose in the case of exposure to radiation in a large number of people In this study, we tried to measure the expression of pg3 and gdfl5 in human peripheral blood lymphocytes induced by ionizing radiation by using the technology of real-time PCR, ELISA and so on. This will help the molecular level biological effect of ionizing radiation and its mechanism, provide a new biological marker for the research of radiation damage, and will also find a more effective, rapid and high-flux biological dosimeter accumulation theory. It was reported. Method:1. Under the condition of informed consent,73 normal persons were collected. quantitative analysis of the expression of pg3 and gdf15 in peripheral blood lymphocytes from peripheral blood samples by using the Taqman MGB probe method the level of the pg3 and gdf15 genes in the peripheral blood lymphocytes of the irradiated human were analyzed by using the Taqman MGB probe method and the ELISA technique, and the dose effect relationship between the transcription level and the translation level of the human peripheral blood lymphocytes after the irradiation (4h, 8h, 12h, 24h, 48h and 72h) was analyzed, and the dose was 0,1, 3,5 and 8 Gy, and the dose response curves were finally fitted by a statistical method; and 3. By analyzing the expression of gdf15 gene in the isolated and whole peripheral blood of SD rats exposed to different doses (OGy, 2Gy, 4Gy. 6Gy), the expression of gdf15 gene in the isolated and whole peripheral blood of SD rats was changed. dosimeter The results were as follows:1. The background copy number of the expression of the pg3 and gdf15 genes in the peripheral blood of the normal human is different from the different sex and not in the normal human. There was no statistical difference between the same age groups. The expression levels of the pg3 and gdf15 gene in the peripheral blood lymphocytes of the human peripheral blood lymphocytes from 8 h to 48 hours after the irradiation of 2.0 to 8 Gy 60Co showed a good dose-effect relationship and can be fitted into a straight line with a higher R2 value. The dose-increasing, the pige3 and the gdf occurred at 72 h after the irradiation. The expression of PIG3 protein in peripheral blood of human peripheral blood showed a dose-effect relationship at 48 h and 72 h after the level of translation, and the expression of PIG3 protein in human peripheral blood lymphocytes could be fit into a straight line with high R2 value. ; others Conclusion:1. The level of the normal human peripheral blood pige3 and gdf15 gene mRNA is not affected by the sex and age of the donor. The expression level of the two genes was the basis of the radiation biological dosimeter.2. The expression of the human peripheral blood pige3 and gdf15 in the in-vitro irradiation at the mRNA level showed a good dose-effect relationship between 8 and 48 hours after the dose of 0-8Gy. The expression of the two genes is the basic condition of the biological dosimeter.3, the level of the PIG3 protein is 0-5.
【学位授予单位】：中国疾病预防控制中心
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R142
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